Product Description

The ProteoSpin™ Detergent Clean-Up Maxi Kit removes up to 99.99% of detergents from protein samples while maintaining high protein recovery. The kit employs spin column technology that offers convenience, speed, and ease-of-use.All types of detergents including ionic, non-ionic and zwitterionic detergents can be effeciently removed in a very short time, eliminating the need for for labourious procedures such as dialysis.The kit is designed to remove detergents in protein solutions either in their free form or bound form, as when complexed with the protein. Also, unlike other commercially available kits that can only handle one detergent and often do notremove denaturants, the ProteoSpin™ Detergent Clean-Up Micro Kit removes a variety of detergents and denturants that may be used during the purification of the protein samplewhile providing high protein recovery.

Breifly, pH Binding Buffer is added to the protein sample and the sample is applied to Norgen's column(BIND). Under these conditions the proteins will bind to the column whiledetergents and other contaminants are removed in the flowthrough. The bound proteins are then washed to remove any remaining impurities (WASH). Lastly, the clean proteins are eluted into a small volume of the provided Elution Buffer or into other optional elution buffers (user-provided) (ELUTE). Please see the procedure flowchart to the right.

No further cleaning of the protein sample is required and can be used directly in a variety of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, and NMR spectroscopy. The kit employs a proprietary matrix, which acts as an ion exchanger This kit offers an additional advantage in that basic proteins can be factionated separately if the pI of the protein is known to be above pH 7.This gives further purification from other contaminating acidic proteins.

FEATURES AND BENEFITS

Fast and easy processing- In contrast to longer procedures such as dialysis, this easy-to-use procedure allows for the processing of4 samples in only 40 minutes.

No detergent carryover - Each sample is processed individually using its own spin column, resulting in no carryover or column bleed from sample to sample.

Efficient detergent removal - Removes more than 95% of detergents, including detergents that are complexed (bound) with proteins

Detergent-free samples are ready to use - No need for additional sample preparation procedures for downstream applications

Ready-to-use columns - No purging, swelling of resin, or any other lengthy preactivation

Specifications

The purified proteins are suitable for a number of downstream applications such as:

2D (Two Dimensional)SDS-PAGE

Whole protein mass spectrometry

Isoelectric focusing

X-ray crystallography

NMR spectroscopy

Protein microarrays.

Table 1. Efficient Removal of SDS.

Removal of SDS with the ProteoSpinTM Detergent Clean-Up Maxi Kit is greater than 99%. To evaluate this, 2 mg samples of BSAwere spiked with 0.1% SDSand 1% SDS, and were processed using the kit. The protocol was followed, and the proteins were bound, washed and eluted as described. The purified protein was then assayed for SDS using the Detergent-OUT kit by Geno Technologies, Inc, and SDSremoval was found to be greater than 99%, with up to 50 mg of SDSbeing removed.

Table 2: High Recovery of Proteins from an SDS-Containing Solution.

The recovery of both acidic and basic proteins from solutions containing SDS is high using the ProteoSpinTM Detergent Clean-Up Maxi Kit. To evaluate recovery, 10 mL buffer samples were spiked withlysozyme or BSA as well as either 0.1% SDS or 1.0% SDSasshown in the table above. The sampleswere processed using the kit as per the provided protocol to remove detergents, and protein recovery was assessed. Recoveries of up to 100%were observed, indicating that high protein recoveriescan be achieved even in the presence of high amounts of detergents.

Storage Conditions
Unopened solutions should be stored at room temperature. Once opened, the solutions should be stored at 4°C when not in use, except for the Basic and Acidic Binding Buffers, which should be stored at room temperature.